A new chemical, Peldri II, is evaluated as a compound for drying soft clay materials. Peldri II, a fluorocarbon (1,1-difluorotetrachloroethane), is a solid at room temperature and is a liquid above 25°C. Clay gels are embedded in Peldri II by immersing them in the liquid and allowing it to solidify. Once solidified, Peldri II will sublime, with or without vacuum, to a dry specimen, probably without introducing surface tension. Wyoming montmorillonite saturated in 10−3 M NaCl and 1 M CaCl2 solutions has been examined to compare preservation of its initial structure after Peldri II, critical point, and freeze drying techniques. Transmission electron microscopy of ultrathin sections, scanning electron microscopy, and mercury intrusion porosimetry techniques were used. No differences were detected between Peldri II and critical point drying methods. Peldri II appears to be a significant improvement as an alternative drying agent for clay materials in studies of their structural properties. It is also very convenient for drying large numbers of samples.

To study the physico-chemical activity of lichens on micaceous components of granitic rocks, samples covered by thalli of Parmelia conspersa (Ehrht) Ach. and Aspicilia intermutans (Nyl.) Arn. were collected and examined with Scanning Electron Microscopy (SEM) equipped with a Back Scattered Electron (BSE) detector and an Energy Dispersive Spectroscopy (EDS) microanalytical system. The bio-physical activity of both lichen species leads to a deep alteration of biotite, which results in detachment, separation and exfoliation of biotite plates. Chemically, the bioweathering process of biotite in the lichenmineral contact zone involves considerable depletion of potassium (K) from interlayer positions in biotite and removal of several elements, corresponding to a 9.7% loss in matter. The sequence of the loss of elements is: K+ » Fetot > Ti4+ ≅ Mg2+. There are also some gains in the order: Ca2+ > Na+ » Al3+ > Si4+ attributed to dissolution of co-existing Ca and Na rich minerals. Geochemical mass balance results suggest the transformation of K-rich biotite to scarcely altered biotite interstratified with a biotite-vermiculite intermediate phase in the lichen bioweathered contact zones.

The question of whether clay minerals can be biogenically transformed as a result of lichen activity at the lichen-rock interface remains unresolved. We applied several microscopical and analytical techniques—scanning electron microscopy-back-scattered electron (SEM-BSE), energy dispersive spectroscopy (EDS) and high-resolution transmission electron microscopy (HRTEM)—in an attempt to address this issue. Unaffected granitic biotite and bioweathered material from the granitic biotite and Parmelia conspersa lichen thalli interface were examined using HRTEM after ultrathin sectioning. The n-alkylammonium treatment of ultrathin sections was carried out in order to study the biogenous mineralogical transformation of the biotite. Microsamples proceeding from unaffected biotite zones demonstrated homogenous 10-Å d(001)-value biotite phase. HRTEM images of lattice fringes of samples taken from the lichen-biotite contact zone reveal large areas of both unexpanded (10-A) and randomly and R = 3 distributed expanded (from 14- to 30-Å) layers of phyllosilicates identified as interstratified biotite-vermiculite. Results of artificial biotite weathering (replacement of K by Ca ion) also revealed the biotite-vermiculite phase formation, indicating that K release in biotite is one of the mechanisms responsible for interstratified mineral phase formation. Two parallel processes, physical exfoliation of biotite and inter-layer ionic exchange of K and subsequent vermiculite formation, are the mechanisms for biotite bioweathering induced by lichens.

Amber is a superb medium for the fossilization of delicate organisms. Besides light microscopy techniques for the study of insects in amber, scanning electron microscopy (SEM) in backscattered electron mode (SEM-BSE), low temperature SEM (LTSEM) and also confocal laser scanning microscopy (CLSM) were used to examine microcenosis and particulate plant remains (microdebris). We applied these techniques to such inclusions in amber Álava, northern Spain (Allaian: Early Cretaceous). Confocal microscopy provides a 3D image of partial microcenosis showing bifurcate fungal hyphae. The huge potential of SEM-BSE yields high resolution images, in which the relationship between protozoa and fungal hyphae may be observed and the characterization of further ultrastructural details in flagellates. According to the SEM-BSE images, food and pulsatile vacuoles appear better preserved than mitochondria and lipids in amber-embedded protozoa. A process of protozoan mineralization has led to the deposition of S and Fe in peripheral areas, and the Fe is also present in the core of surrounding fungal hyphae. Application of LTSEM for the study of protozoan inclusions produces images of their exteriors showing many vacuoles. Plant tissues under SEM-BSE show mummified cell walls, while the cytoplasm exhibits a bright appearance and is very rich in Fe and S. SEM in secondary electron mode (SEM-SE) also reveals a microbiota trapped in gas bubbles.

We review the lithic microbial ecosystems of the McMurdo Dry Valleys as the main form of terrestrial colonization in this region, and assess the role of environmental controls such as temperature, solar radiation, water availability, wind, nutrient availability, salinity and the physicochemical properties of the colonized rock. Epilithic communities, especially those dominated by lichens, are able to withstand extreme environmental conditions but subsurface endolithic microhabitats provide more tolerant conditions. Endolithic microbial communities can be grouped into two main classes: eukaryotic communities (dominated by lichenized fungi and algae) and prokaryotic communities (dominated by cyanobacteria). Heterotrophic bacteria and non-lichenized algae and fungi (mainly black fungi) are also components of these communities. These lithobiontic microorganisms generally have effective mechanisms against freezing temperatures and desiccation. Extracellular polymeric substances play an important role not only in protecting microbial cells but also in community organization and in mitigating microenvironmental conditions. Antarctic lithobiontic communities are comprised of microbial consortia within which multiple interactions between the different biological and abiotic components are essential for microbial survival, whilst fossils and biomarkers provide evidence of earlier successful microbial life in Antarctic deserts. Finally, the uniqueness of the present lithobiont assemblages suggests they are the outcome of geographical isolation during the evolution of the continent and not merely the descendants of a subset of globally distributed taxa that have adapted to the extreme environmental conditions.

In this study, the symbiont cells of several endolithic lichens colonizing granite in continental Antarctica and the relationships they have with the abiotic environment were analyzed in situ, in order to characterize the microecosystems integrating these lichens, from a microecological perspective. Mycobiont and photobiont cells, the majority classified as living by fluorescent vitality testing, were observed distributed through the fissures of the granite. The fact that extracellular polymeric substances were commonly observed close to these cells and the features of these compounds, suggest a certain protective role for these substances against the harsh environmental conditions. Different chemical, physical and biological relationships take place within the endolithic biofilms where the lichens are found, possibly affecting the survival and distribution of these organisms. The alteration of bedrock minerals and synthesis of biominerals in the proximity of these lichens give rise to different chemical microenvironments and suggest their participation in mineral nutrient cycling.

The effects of the structural features of the thallus and the lichen–substratum interface on soil water infiltration were investigated using low temperature scanning electron microscopy and scanning electron microscopy with backscattered electron imaging. Two morphologically different lichen species were examined, the crustose Diploschistes diacapsis and the squamulose Squamarina lentigera. Water distribution within lichen-forming soil crusts explains the results of earlier infiltration studies. Several anatomical features of the upper cortex gave rise to similar impervious characteristics of the thallus. The fungal material comprising the lichen thallus showed significant hydrophobic features and only when in contact with calcium oxalate crystals or minerals in the soil was this material able to retain water. The lichen–substratum interface appears to play an important role in the hydric strategy of these lichens which in turn affects the penetration of water into the soil.

Rimularia insularis forms an photosynthetic thallus on lichens of the Lecanora rupicola group. In the thalli of R. insularis, some hyphae of the host are detectable, and they are situated mainly in the basal part of the Rimularia's medulla. No Lecanora hyphae were present in the Rimularia's algal layer and they were indistinct in the upper parts of its medulla, from where we obtained clean ITS-sequences of Rimularia. Direct PCR from various sections of the algal layer detected only one photobiont genotype, although the algal cells could be assigned to two morphologically different types. Calcium deposits were found in the upper parts of the medulla, while abundant rock substrate fragments with different mineral compositions were present in the lower parts. Cavities and fissures in the rock below the infected thalli were filled by fungal cells, PCR analyses of such parts indicated that the Rimularia hyphae extend below the host, and into the rock.

Cryptoendolithic microbial communities living within Antarctic rocks are an example of survival in an extremely cold and dry environment. The extinction of these micro-organisms formerly colonizing sandstone in the Mount Fleming area (Ross Desert), was probably provoked by the hostile environment. This is considered to be a good terrestrial analogue of the first stage of the disappearance of possible life on early Mars. To date, only macroscopically observed indirect biomarkers of the past activity of cryptoendoliths in Antarctic rocks have been described. The present paper confirms, for the first time, the existence of cryptoendolith microbial fossils within these sandstone rocks. The novel in situ application of scanning electron microscopy with backscattered electron imaging and simultaneous use of X-ray energy dispersive spectroscopy allowed the clear detection of microfossils left behind by Antarctic endoliths. Careful interpretation of the morphological features of cells, such as preserved cell walls in algae, fungi and bacteria, cytoplasm elements such as chloroplast membranes in algae and organic matter traces, mineral associations, and the spatial context of these structures all point to their identification as cryptoendolith microfossils. This type of investigation will prompt the development of research strategies aimed at locating and identifying the signs that Martian microbiota, probably only bacteria if they existed, may have been left for us to see.

The chemical and ultrastructural features of the interface formed by different biotypes of saxicolous lichen species with their rock substrata were investigated in the semiarid habitat of the SE Iberian Peninsula and the relationships between the bioweathering patterns observed and lichen colonization selectivity towards the different rock substrata evaluated. Xanthoria parietina was able to fix to the rock substratum by the adherence of single cell walls from the lower cortex. Neofuscelia pulla used rhizines and loose groups of hyphae for attachment of the thallus to the rock. Colonization by the foliose lichen species was confined to the rock surface, while Diploschistes diacapsis was also able grow below the surface showing two types of hyphal growth. Minerals in close contact with cell walls were biochemically and biophysically weathered, but hyphae showing calcium oxalate crystals did not appear to be directly involved in the patterns observed. The textural characteristics of the substratum seemed to be related to the type of microorganism colonization: sedimentary rocks were more deeply colonized by lichens and other chasmolithic microorganisms than volcanic material. Calcium oxalate crystals were found in the medulla of N. pulla but not at the lichen-substratum interface. Crustose lichens such as D. diacapsis showed calcium oxalate crystals in the upper cortex and over the outside of fungal medullary hyphae but not in direct contact with the rock surface. Calcium oxalate precipitation seems to be related to the different metabolic activities of the mycobiont within the lichen thallus and to different species. D. diacapsis inhibits the growth of other microorganisms in close proximity to the thallus, whereas foliose species were associated with several communities of microorganisms.

Linear mycelial aggregates (rhizomorphs) produced by the squamulose lichens Toninia opuntioides, Squamarina cartilaginea, Lecanora rhizinata, and Acarospora scotica were examined by transmission electron microscopy (TEM). Cytological features typical of lichenized mycobionts were present, including concentric bodies and plasmalemma invaginations. Some septa in rhizomorphs of S. cartilaginea and A. scotica appeared to be multipored. Septal pores were usually plugged by an electron-dense structure. Intrahyphal hyphae were common to abundant in the rhizomorphs of all four species examined. The wall of the invasive hypha was continuous with inner wall layers of the cell of origin. Contacts between adjacent hyphae involved wall dissolution and formation of new wall layers, leading to anastomosis and/or possibly to intrahyphal invasion. Differentiation of an outer layer of collapsed cells in rhizomorphs of Squamarina cartilaginea and of extremely thick-walled cells Lecanora rhizinata was evident, but no anatomical specialization comparable to the ‘vessel hyphae’ of non-lichen linear mycelial organs was observed. Enlarged spheroid or ovoid cells with dense cytoplasm, numerous mitochondria and lipid reserves occurred in short chains in rhizomorphs of Acarospora scotica; infrequently, similar cells were observed singly in those of Squamarina cartilaginea. These cells were reminiscent in shape of certain types of oil hyphae found in endolithic lichens, although often the lipid content of the enlarged cells was not particularly pronounced.